EP2404509A1 - Traubenkernextrakte, Verfahren zu ihrer Herstellung und Verwendung dafür zur Behandlung von Wein - Google Patents

Traubenkernextrakte, Verfahren zu ihrer Herstellung und Verwendung dafür zur Behandlung von Wein Download PDF

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Publication number
EP2404509A1
EP2404509A1 EP11173214A EP11173214A EP2404509A1 EP 2404509 A1 EP2404509 A1 EP 2404509A1 EP 11173214 A EP11173214 A EP 11173214A EP 11173214 A EP11173214 A EP 11173214A EP 2404509 A1 EP2404509 A1 EP 2404509A1
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Prior art keywords
extraction
grape seed
wine
grape
protein
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English (en)
French (fr)
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Andrea Curioni
Simone Vincenzi
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Universita degli Studi di Padova
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Universita degli Studi di Padova
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12H—PASTEURISATION, STERILISATION, PRESERVATION, PURIFICATION, CLARIFICATION OR AGEING OF ALCOHOLIC BEVERAGES; METHODS FOR ALTERING THE ALCOHOL CONTENT OF FERMENTED SOLUTIONS OR ALCOHOLIC BEVERAGES
    • C12H1/00—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages
    • C12H1/02—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages combined with removal of precipitate or added materials, e.g. adsorption material
    • C12H1/04—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages combined with removal of precipitate or added materials, e.g. adsorption material with the aid of ion-exchange material or inert clarification material, e.g. adsorption material
    • C12H1/0416—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages combined with removal of precipitate or added materials, e.g. adsorption material with the aid of ion-exchange material or inert clarification material, e.g. adsorption material with the aid of organic added material
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
    • A23L11/30—Removing undesirable substances, e.g. bitter substances
    • A23L11/32—Removing undesirable substances, e.g. bitter substances by extraction with solvents
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/185—Vegetable proteins
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12F—RECOVERY OF BY-PRODUCTS OF FERMENTED SOLUTIONS; DENATURED ALCOHOL; PREPARATION THEREOF
    • C12F3/00—Recovery of by-products
    • C12F3/06—Recovery of by-products from beer and wine
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

  • the invention relates to grape seed extracts usable in the oenological field for improving the organoleptic characteristics and for clarification/stabilization of wines and to a method of preparing said extracts.
  • the invention further relates to the use of grape seed extracts for organoleptic improvement and clarification/stabilization of wines by means of a protein fining agent.
  • the first approach when assessing a wine includes among its parameters the degree of clarity, which depends on the presence or absence of particles in suspension.
  • degree of clarity which depends on the presence or absence of particles in suspension.
  • Wine is a colloidal system, in which the colloids are represented by proteins, polysaccharides, protein-polyphenol and protein-polysaccharide aggregates.
  • a colloidal solution consists of particles that are maintained in dispersion in a liquid by a number of forces, including electrostatic repulsion and hydrophilicity. The latter provides a hydration layer around the macromolecule which helps to keep it suspended. Under the action of various factors, however, the (invisible) colloidal particles can aggregate (flocculation), forming particles that are visible as turbidity, which can eventually settle to the bottom of the bottle.
  • a treatment is applied to the wine (called protein fining) which causes the early removal, by precipitation, of the unstable colloids, obtaining both a clarifying and a stabilizing effect.
  • the technique comprises adding, to a wine with a certain degree of turbidity (already present and/or potential), a proteinaceous substance that is able to flocculate and settle, entraining with it the particles responsible for any turbidity that is already present, but also those that are potentially able to cause clouding at a later date.
  • the insoluble material resulting from the treatment is then easily removed from the wine.
  • the list also includes eggs and egg-based products, milk and milk-based products, cereals containing gluten, soya and soya-based products, lupins and lupin-based products, all of which are products that must be declared on labels of food products, including wine.
  • the proteinaceous oenological aids used in fining of wine should not, in practice, remain in the finished product, it is important to bear in mind that the presence of residues of said proteins cannot be ruled out, thus representing a potential risk to the health of consumers who are sensitive to the raw material from which the oenological clarifiers were extracted.
  • the aim of the present invention is to provide a protein product, of homologous origin relative to the wine itself, for use in the fining of wines, thus avoiding recourse to materials extraneous to grapes.
  • the inventors have identified an extract derived from vegetable materials homologous to the product to be treated, namely wine, having the necessary characteristics for use as oenological aid in the fining of wine and have developed a method of preparation suitable for obtaining said extract.
  • the extract must have a minimum protein content, such as to produce an effective fining process, and at the same time should have a content of polyphenols that does not affect the organoleptic properties of the wine treated or to be treated.
  • compositions comprising grape seed extracts, having a content of protein nitrogen (measured as mineral nitrogen multiplied by a conversion factor of 6.25) in an amount higher than at least 30% (w/w) and polyphenols (determined with Folin-Ciocalteu reagent) in an amount lower than 40% (w/w).
  • the present invention relates to a method of preparation of compositions comprising grape seed extracts, having the characteristics stated above, comprising at least the steps of:
  • the starting materials can be either fresh grape seeds or residues from extraction of grape seed oil.
  • the materials on which extraction is performed consist of fresh grape seeds, these are subjected to a step of grinding and delipidation by treatment with an organic solvent before extraction.
  • the solid residue obtained from the first extraction can be removed or submitted, as it is or combined with new material from grape seeds, to one or more further extractions.
  • the protein precipitate obtained by acidification can be purified by subsequent dialysis.
  • a key problem is to obtain a product that has a minimum content of proteins so as to produce efficient clarification of the wine treated without causing organoleptic changes of the latter due to a high content of polyphenols, which are difficult to remove from said extract.
  • the first technical aspect to be solved is on which type of material to carry out extraction, being this one the key step of the entire process of preparation of the extract.
  • the vegetable material can be either fresh material (grape seeds) or processing residues from extraction of oil (so-called extraction flour).
  • extraction flour processing residues from extraction of oil
  • the flours obtained after industrial extraction of oil from grape seeds, already being by their nature free from lipids are to be preferred.
  • organic solvents suitable for extracting the lipids are selected from hexane and petroleum ether.
  • the main parameter to be taken into account for extraction of the proteins from the delipidated vegetable material is the pH.
  • aqueous solution that can also be constituted from water alkalized with a base (NaOH) or from a suitable buffer (for example Glycine buffer, sodium carbonate buffer or Tris buffer) at concentrations between 0.05 and 0.2 M.
  • a suitable buffer for example Glycine buffer, sodium carbonate buffer or Tris buffer
  • the extraction condition that has been shown to result in obtaining an extract suitable for the use envisaged is related to pH of the aqueous extraction solution, being the pH 10.5 the most effective, while the composition of the extractant or its molarity (in the range between 0.05 and 0.2 M) were not found to have a significant influence on the quality of the extract.
  • Extraction must be carried out with a solid:liquid ratio of at least 1:5 and for a time of at least 3 hours.
  • a solid:liquid ratio of 1:10 By keeping the pH constant throughout the extraction phase and using a solid:liquid ratio of 1:10, it is possible to extract more than 80% of the protein material in just the first 3 hours of extraction.
  • the solid residue obtained from extraction is then separated from the liquid phase by filtration or centrifugation or with other equivalent means.
  • This solid residue can be submitted to a new extraction, in conditions similar to the first, and preferably for a period of time between 10 and 12 hours.
  • the liquid phase separated by extraction is acidified to pH 3.0 by adding 6N hydrochloric acid.
  • 6N hydrochloric acid At this pH, some of the proteins present in the extract precipitate (Fazio et al., 1983, ref. cit. ; El-Aal, M.H.A, 1992, Die Exercise 2: 112-118 ).
  • the protein precipitate can be recovered by centrifugation or by decanting.
  • the precipitate, resuspended in slightly basic solution (0.01 N NaOH) to redissolve the proteins, is subsequently submitted to a step of purification by dialysis against water.
  • the contents of the dialysis tube are then removed and lyophilized.
  • an extract is obtained with a content of protein nitrogen (total nitrogen, determined by mineralization by Kjeldahl's method and converted to protein nitrogen by multiplying by a factor of 6.25) equal to about a third of the total weight of the preparation (30-38%).
  • total nitrogen determined by mineralization by Kjeldahl's method and converted to protein nitrogen by multiplying by a factor of 6.25
  • the content of total polyphenols determined by colorimetric analysis with Folin-Ciocalteu reagent ( Singleton, V.L. and Rossi, J.A., 1965 Am. J. Enol. Vitic. 16:144-158 ) was found to be rather high (33-38% of the total weight of the preparation).
  • the presence of such a high level of polyphenols in the preparations does not constitute a problem from the organoleptic standpoint, since chemical and sensory analyses have not indicated release of polyphenols in the wine by protein preparations from grape seeds. Owing to the high content of polyphenols, the lyophilized extract has a deep reddish-brown coloration, but when tasted, there is no sensation of astringency, or of bitter or sour taste.
  • the dry protein extract prepared as described above, can be taken up in 0.01 N NaOH.
  • a solution of grape seed proteins is obtained which, when added to the wine to be treated in an amount between 20 and 40 g/hL, makes it possible to obtain technological performance similar to that of the animal gelatin normally used in the fining of red wines.
  • Sensory analysis of the wines treated has also demonstrated similar effects to those obtainable using gelatin, as will be clear from the description given below of the tests carried out.
  • the chemical, nephelometric and sensory analyses used for evaluating the effects of the preparations obtained on the characteristics of a red wine are described in detail below.
  • composition in the grape seed extracts obtained with the two different buffers used glycine and sodium carbonate buffers
  • polyphenolic fraction based on the ratio of the absorbances at 280 and at 420 nm
  • the main parameter taken into consideration for extraction of the proteins was the pH.
  • the first extraction was carried out for 3 hours with a solid:liquid ratio of 1:10.
  • the residue of the first extraction was extracted further, overnight, with fresh buffer in order to obtain exhaustive extraction from the matrix.
  • the solid residue was separated by centrifugation (5 minutes at 3000g).
  • the supernatants obtained were filtered (0.45 ⁇ m), and were of a deep colour.
  • the final pH of the liquid extracts obtained was measured (Table 1). Table 1.
  • the pH of the extract found was, except in the case of extraction with Tris-HCl buffer, lower than the pH of the buffer solution used for extraction.
  • the absorbance at 280, 320 and 420 nm was then determined on all the extracts.
  • the extracts in glycine buffer (indicated by the ovals in Fig. 1 ) always showed greater absorbance than that of the extracts in sodium carbonate.
  • the ratio 280/420 nm is particularly discriminating, and was found to be very high for the extracts in glycine buffer, with a decrease with increase in pH. In contrast, the same ratio is very low for extracts in sodium carbonate buffer. Considering that the total absorbance at 280 nm is very similar for the two buffers ( Fig. 1 ), this would indicate that sodium carbonate extracts a larger amount of brown substances (probably polyphenols) which absorb at 420 nm.
  • the yields, relative to the initial weight of flours extracted came to about 3-4% (w/w) in extractions at pH 10.5 regardless of the buffer used. Then it was verified that, taking care to correct the pH continuously with 1 M NaOH to counter the acidifying effect of the grape seed flours, the same yield can also be obtained with a single extraction of just 3 hours.
  • the grape seed proteins have minimum solubility at pH 3.0, this property was utilized for precipitating them from the extract, at the same time removing all the proteins potentially soluble at the pH of the wine.
  • the extracts were, therefore, treated with 6N HCl, lowering the pH to 3.0.
  • the resultant precipitate was recovered by centrifugation (10 min at 3000g), taken up in water, and dialysed exhaustively against water using membranes with cut-off of 3.5 kDa. The contents of the dialysis tube were then removed and lyophilized. The weight of the lyophilized powder was used for calculating the yield (weight/weight) of the preparation process.
  • the lyophilized preparations (precipitates obtained from the extracts by precipitation at pH 3.0) were dissolved at 40 mg/ml in 0.01 N NaOH. The solutions thus obtained were then added to an unclarified red wine (Merlot, 2009 vintage) to obtain final concentrations of 20 and 40 g of lyophilizate/hL of wine. The same wine was also treated with commercial oenological gelatin at 20 g/hL. The samples were left to decant at 20°C for 48 h and after this time the samples of wine were centrifuged at 1900g x 10 minutes (Maury et al., 2003, ref.
  • the extracts in glycine have greater clarifying capacity than the extracts in sodium carbonate. The same result was also obtained on other red wines. Moreover, the grape seed extracts obtained with glycine have an effect of reduction of turbidity entirely comparable to that of commercial gelatin, showing that they are effective in the process of clarification of red wine.
  • the instrumental index AMI (Astringency Mucin Index) was used, which is correlated linearly with the astringency perceived by a group of trained tasters ( Monteleone E. et al., 2004, Food Quality and Preference 15: 219-227 ).
  • the index is determined using an in vitro test designed according to the principles of predictive models of the simulation type. Mucin is used as model salivary protein. The ratio of the model saliva/stimulus volumes, the reaction pH and physiological temperature of 37°C reproduce the conditions normally prevailing in the mouth. Interaction between polyphenols and mucin and the formation of insoluble aggregates leads to the generation of turbidity, which is measured nephelometrically and expressed in nephelometric turbidity units (NTU).
  • a commercial extract of tannins from grape seeds (Graptan PC, Inntec, Verona) was used, at 1.8 mg/ml in 1 % ethanol, as a model phenolic system.
  • results of the test of Least Significant Difference (LSD) indicate that the AMI values of solutions of tannins treated with the preparations from grape seeds are significantly lower than that of the untreated solution of tannins ( Fig. 3 ).
  • treatment with commercial oenological gelatin was found to be the most effective in reducing the AMI value of the solution of tannins.
  • the model solutions of tannins treated with the preparations from grape seeds and with commercial oenological gelatin were assessed by a group of trained tasters, who were asked to assess bitterness, acidity and astringency and to express the perceived intensity on the assessment scale (Labelled Magnitude Scale).
  • the protein preparations from grape seeds produced a significant decrease of astringency, although to a lower extent than oenological gelatin.
  • sensory analysis did not find significant differences between the various protein preparations from grape seeds.
  • the AMI values of the polyphenols extracted from the treated wines were submitted to one-way analysis of variance to assess the effect of the treatment on removal of potentially astringent polyphenolic components.
  • the results of the LSD test indicate that the treatment with the protein preparations from grape seeds gives a significant reduction in the AMI value of the phenolic extracts of the wines relative to the control (untreated wine), but do not demonstrate significant differences for the various preparations.
  • oenological gelatin is confirmed as the most effective aid for removing potentially astringent phenols ( Fig. 5 ).
  • the treated and untreated wines were finally submitted to sensory analysis by two different panels: one specifically trained for the perception of bitterness, acidity and astringency (panel 1) and a panel of tasters experienced in assessment of the overall quality of wines (panel 2).
  • the scores obtained by the trained panel (panel 1) were submitted independently to one-way analysis of variance for evaluating the effect of the treatments on the sensory properties relating to the intensity of bitterness, of acidity and of astringency of the wine.
  • the results demonstrated that treatment with the preparations does not cause any significant difference in intensity of bitterness and acidity (as already observed with the model solution of grape seed tannins) ( Figs. 6 and 7 ).
  • no significant differences were found in perceived astringency in the wines treated with the protein preparations from grape seeds relative to that treated with oenological gelatin.
  • the panel of expert tasters assessed the wines with a points-based test using a specific unstructured rating scale for Cabernet Sauvignon.
  • the results relating to astringency and to the overall assessment were processed separately. Again in this case the scores were submitted independently to one-way analysis of variance.
  • the results confirmed that the astringency of the wines was perceived in a manner significantly different between the treatments (F 7.65; p ⁇ 0.0001) ( Fig. 9 ).
  • the results of the LSD test showed that in the treated wines, both with gelatin and with protein preparations from grape seeds, the perception of astringency was lower than for the untreated wine, whereas no differences were found between the different treatments.
  • the "radar screens" in Fig. 11 show curves corresponding to the two different tasting sessions.
  • the only statistically significant differences were the improvement of olfactory harmony and the decrease in acidity in the wine treated with gelatin relative to the control (the decrease in acidity observed in the wine treated with glycine is not significant).
  • Comparison of the effects of the various protein preparations from grape seeds did not find significant differences for any of the descriptors.
  • gelatin by removing a good part of the anthocyanins (12.5%), can cause an excessive reduction in red colour, mainly important for those wines that already have a medium-low content of anthocyanins (for example Pinot Noir, Valpolicella, etc.).
  • the grape seed preparations have a smaller effect on total anthocyanins (Table 4).
  • the preparations from grape seeds also contribute to their oxidative stability.
  • the test of oxidizability of anthocyanins measured as resistance to bleaching by hydrogen peroxide, demonstrated that the wines treated with the preparations from grape seeds have an oxidizability index equal to or even less than that of the wine treated with gelatin.

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  • Engineering & Computer Science (AREA)
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  • Food Science & Technology (AREA)
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  • Proteomics, Peptides & Aminoacids (AREA)
  • Agronomy & Crop Science (AREA)
  • Distillation Of Fermentation Liquor, Processing Of Alcohols, Vinegar And Beer (AREA)
  • Peptides Or Proteins (AREA)
EP11173214A 2010-07-08 2011-07-08 Traubenkernextrakte, Verfahren zu ihrer Herstellung und Verwendung dafür zur Behandlung von Wein Withdrawn EP2404509A1 (de)

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ITPD2010A000214A IT1400808B1 (it) 2010-07-08 2010-07-08 Estratti da vinaccioli, metodo di preparazione e uso degli stessi per il trattamento di vini.

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018178119A1 (de) 2017-03-28 2018-10-04 Dietz Max Verfahren zur prozessökonomischen ab-/auftrennung von konstituenten pflanzlicher ausgangsmaterialien sowie deren gewinnung und verwendung
EP3498103A1 (de) * 2017-12-13 2019-06-19 Drei Lilien Pvg GmbH&Co. KG Verfahren zur prozessökonomischen ab-/auftrennung von konstituenten pflanzlicher ausgangsmaterialien sowie deren gewinnung und verwendung
CN115232689A (zh) * 2022-06-22 2022-10-25 茅台学院 一种葡萄酒泥废弃物全组分利用的生物炼制方法
CN118599619A (zh) * 2024-05-29 2024-09-06 茅台学院 一种基于酒糟蛋白制作澄清剂的制造方法

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FR361125A (fr) * 1905-12-28 1906-05-18 Georges Eugene Jacquemin Procédé d'extraction d'une substance azoto-phosphorée contenue dans les résidus de raisins en particulier et dans les végétaux de consommation ou leurs résidus en général
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Cited By (8)

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CN110475478A (zh) * 2017-03-28 2019-11-19 马克斯·迪茨 切断/分级分离植物起始材料中成分的工艺经济的方法及其生产和用途
RU2767338C2 (ru) * 2017-03-28 2022-03-17 Макс ДИТЦ Способ отделения/разделения компонентов растительного сырья, а также их извлечение
CN110475478B (zh) * 2017-03-28 2024-01-02 马克斯·迪茨 切断/分级分离植物起始材料中成分的工艺经济的方法及其生产和用途
US11856968B2 (en) 2017-03-28 2024-01-02 Max Dietz Method for economical removal/fractionation of constituents of vegetal starting materials
EP3498103A1 (de) * 2017-12-13 2019-06-19 Drei Lilien Pvg GmbH&Co. KG Verfahren zur prozessökonomischen ab-/auftrennung von konstituenten pflanzlicher ausgangsmaterialien sowie deren gewinnung und verwendung
CN115232689A (zh) * 2022-06-22 2022-10-25 茅台学院 一种葡萄酒泥废弃物全组分利用的生物炼制方法
CN118599619A (zh) * 2024-05-29 2024-09-06 茅台学院 一种基于酒糟蛋白制作澄清剂的制造方法

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